Preparation And Identification Of Monoclonal Antibodies Against 53ku ES Recombinant Protein Of Trichinella Spiralis

Trichinellosis was a zoonotic parasitic disease caused by Trichinella spiralis. It mainly parasitized in more than hundred and fifty kinds of animals, such as swine,sus scrofa,mice and bear, it also parasitized in people. People ate the pork or the meat of other animals which had Muscle larvae, without cooking or cooking thoroughly to contract the disease. Because of the complexity of the etiological agent spreading, for more than hundred years form discovering the disease, people not only did not completely control it, but also the circumsciption of it was more and more wide. To control and prevent the Trichinellosis quickly and effectually, we must have fast and precise detection technique. The investigation chose P53 ES recombination protein of Trichinella to immunize mice for preparing McAb. McAb will be useful in developing diagnosis kit of Trichinellosis.BALB/c mice were immunized by recombinatnt antigen 53ku of Trichinella spiralis. Their spleen lymph cell were fused with SP2/0 myeloma cell of mice. Masculine hybridoma cell was cloned and sieved by limiting dilution assay and indirect enzyme-linked immunosorbent assay (iELISA), and extracted and purified ascites. Class or subclass of immunoglobulin was examined by the kit. The activity of hybridoma cell strain culture medium and ascites were tested with indirect ELISA. Pecificity of two McAbs were tested by Western-blot. The stability of hybridoma antibody secretion were tested. The ELISA were used to characterize these McAbs by antigens of several parasite including adult antigen of Schistosoma japonicum,Trichuris suis and Cryptosporidium, cyst liquid antigen of Cysticercus cellulosae and coenurus. Indirect immunofluorescence with the monoclonal antibody used to detect muscle larvae immunological reaction. Results indicated:Two hybridoma cell strains secreting the highest titer McAbs were acquired to name 2H5 and 3D2. Both McAbs belonged to IgM subclass withκchain. The titer of 2H5 and 3D2 ascites were 1:12000,l:24000. Westem blot analysis showed that both McAbs reacted with ES (53ku) antigen, The hybridoma cell could grow well and the titer was stabilization after culturing. They had no cross reaction with all mentioned antigens. McAbs and muscle larvae had immunological reactions by Indirect immunofluorescence. The characterized results demonstrated that McAbs would be useful in reaserch of Trichinellosis and developing diagnosis kit of Trichinellosis.